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Brought to light – chromobodies reveal changes in endogenous protein concentration in living cells

21.09.2018

Scientists at the Natural and Medical Sciences Institute (NMI) in Reutlingen and the Eberhard Karls University of Tuebingen have developed new molecular probes to monitor and quantify changes in the concentration of endogenous proteins by live-cell fluorescence microscopy.

In a study now published in Molecular & Cellular Proteomics, Keller et al. describe how fluorescently labeled intrabodies (so-called chromobodies) are stabilized in the presence of their target proteins.


Antigen-Mediated-ChromoBody-Stabilization (AMCBS)

NMI


Antigen-Mediated-ChromoBody-Stabilization (AMCBS)

NMI

Based on this newly uncovered property of chromobodies, the authors present a broadly applicable strategy to optimize chromobodies in order to visualize and measure changes of endogenous target proteins within living cells.

Currently available procedures to measure cellular protein concentration such as quantitative immunoblotting or mass spectrometry analysis are very laborious and time consuming.

Moreover, these approaches require destructive sample preparation and thus only deliver results at a single point in time.

“Our new approach using antigen-mediated chromobody stabilization (AMCBS) allows us to continuously visualize and quantify dynamic changes of specific endogenous proteins directly within living cells – something that could not be done before”, stated Bettina-Maria Keller, the study´s first author.

“We expect this approach to enable unprecedented insights into the dynamic regulation of proteins, e.g. during cellular signaling, cell differentiation, or upon drug action”.
Publication:
Chromobodies to quantify changes of endogenous protein concentration in living cells
Molecular & Cellular Proteomics September 18, 2018, mcp.TIR118.000914; https://doi.org/10.1074/mcp.TIR118.000914

Wissenschaftliche Ansprechpartner:

Prof. Dr. Ulrich Rothbauer
NMI /Universität Tübingen
Markwiesenstrasse 55
72770 Reutlingen
Email: ulrich.rothbauer@nmi.de
ulrich.rothbauer@uni-tuebingen.de
Tel.: +49 (0)7121 30 415

Originalpublikation:

Chromobodies to quantify changes of endogenous protein concentration in living cells
Molecular & Cellular Proteomics September 18, 2018, mcp.TIR118.000914; https://doi.org/10.1074/mcp.TIR118.000914

Weitere Informationen:

http://www.nmi.de

Dr. Nadja Gugeler | idw - Informationsdienst Wissenschaft

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